SUMMARY NOTIFICATION INFORMATION FORMAT (SNIF) FOR PRODUCTS CONTAINING GENETICALLY MODIFIED HIGHER PLANTS (GMHP) Table of contents

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1 SUMMARY NOTIFICATION INFORMATION FORMAT (SNIF) FOR PRODUCTS CONTAINING GENETICALLY MODIFIED HIGHER PLANTS (GMHP) Table of contents A. GENERAL INFORMATION Details of notification...5 (a) Member State of notification... 5 (b) Notification number...5 (c) Name of the product (commercial and other names)...5 (d) Date of acknowledgement of notification Notifier... (a) Name of notifier... 5 (b) Address of notifier... 5 (c) Is the notifier... (d) In case of import the name and address of the manufacturer should be given General description of the product...5 (a) Name of the recipient or parental plant and the intended function of the genetic modification...5 (b) Any specific form in which the product must not be placed on the market (seeds, cut-flowers, vegetative parts, etc.) as a proposed condition of the authorisation applied for...6 (c) Intended use of the product and types of users...6 (d) Any specific instructions and/or recommendations for use, storage and handling, including mandatory restrictions proposed as a condition of the authorisation applied for...6 (e) If applicable, geographical areas within the E.U. to which the product is intended to be confined under the terms of the authorisation applied for...6 ()Any type of environment to which the product is unsuited... 6 (g) Any proposed packaging requirements (h) Any proposed labelling requirements in addition to those required by law... 7 ()Estimated potential demand...7 (j) Unique identification code(s) of the GMO(s) Has the GMIHP referred to in this product been notified under part B of Directive 200 1/18/EC and/or Directive 90/220/EEC? Is the product being simultaneously notified to another Member State? Has the same GMHP been previously notified for marketing in the Community? Measures to take in case of unintended release or misuse as well as measures for disposal and treatment...8 SNIF from insect-protected maize line MJON 863 and maize hybrid MON 863 x MON 810

2 B. NATURE OF THE GMHP CONTAINED IN THE PRODUCT... 9 INFORMATION RELATING TO THE RECIPIENT OR (WHERE APPROPRIATE) PARENTAL PLANTS Complete name...9 (a) Family name...9 (b) Genus... (c) Species... 9 (d) Subspecies... (e) Cultivar/breeding line...9 (f) Common name (a) Information concerning reproduction...9 (b) Sexual compatibility with other cultivated or wild plant species Survivability (a) Ability to form structures for survival or dormancy (b) Specific factors affecting survivability, if any Dissemination (a) Ways and extent of dissemination (b) Specific factors affecting dissemination, if any... I Geographical distribution of the plant...i In the case of plant species not normally grown in the Member State(s), description of the natural habitat of the plant, including informnation on natural predators, parasites, competitors and symbionts I Potentially significant interactions of the plant with other organisms in the ecosystem where it is usually grown, including information on toxic effects on humans, animals and other organisms Phenotypic and genetic traits INFORMATION RELATING TO THE GENETIC MODIFICATION Description of the methods used for the genetic modification Nature and source of the vector used Size, source [name of donor organism(s)] and intended function of each constituent fragment of the region intended for insertion INFORMATION RELATING THE GMHP Description of the trait(s) and characteristics which have been introduced or modified Information on the sequences actually inserted/deleted/modified (a) Size and structure of the insert and methods used for its characterisation, including information on any parts of the vector introduced in the GMHP or any carrier or foreign DNA remaining in the GMHP (b) In case of deletion, size and function of the deleted region(s) I11 SNIF 2 Placing on the market of mnaize grains derived,from insetct-protected maize line MON 863 and maize hybrid MON 863 x MON 81 0

3 (c) Location of the insert in the plant cells (integrated in the chromosome, chloroplasts, mitochondria, or maintained in a non-integrated form), and methods for its determination (d) Copy number and genetic stability of the insert (e) In case of modifications other than insertion or deletion, describe function of the modified genetic material before and after the modification as well as direct changes in expression of genes as a result of the modification Information on the expression of the insert...16 (a) Information on the expression of the insert and methods used for its characterisation (b) Parts of the plant where the insert is expressed (e.g. roots, stem, pollen, etc.) Inform-ation on how the GMHP differs from the recipient plant in...17 (a) Mode(s) and/or rate of reproduction (b) Dissemination (c) Survivability (d) Other differences Potential for transfer of genetic material from the GMHP to other organisms Information on any harmful effects on human health and the environment, arising from the genetic modification Information on the safety of the GMHP to animal health, where the GMHP is intended to be used in animal feedstuffs, if different from that of the recipient/parental organism(s) Mechanism of interaction between the GMHP and target organisms (if applicable), if different from that of the recipient/parental organism(s) Potentially significant interactions with non-target organisms, if different from the recipient or parental organism(s) Description of detection and identification techniques for the GMHP, to distinguish it from the recipient or parental organism(s)...21 INFORMATION ON THE POTENTIAL ENVIRONMENTAL IMPACT FROM THE RELEASE OF THE GMHP Potential environmental impact from the release or the placing on the market of GMOs (Annex II, D2 of Directive 2001/18/EC), if different from a similar release or placing on the market of the recipient or parental organism(s) Potential environmental impact of the interaction between the GMHP and target organisms (if applicable), if different from that of the recipient or parental organism(s) Possible environmental impact resulting from potential interactions wvith nontarget organisms, if different from that of the recipient or parental organism(s). 22 SNIF 3 Placing on the tnarkct of maize grains derived fi-om insect-protected maize line MON 863 and maize hybrid MON 863 x A'!ON 81 0

4 C. INFORMATION RELATED TO PREVIOUS RELEASES History of previous releases notified under Part B of the Directive 2001/18/EC and under Part B of Directive 90/220/EEC by the same notifier...22 (a) Notification number (b) Conclusions of post-release monitoring (c) Results of the release in respect to any risk to human health and the environment (submitted to the Competent Authority according to Article 10 of Directive 200 1/1 8/EC) History of previous releases camred out inside or outside the Community...22 (a) Release country (b) Authority overseeing the release...23 (c) Release site (d) Aim of the release (e) Duration of the release (t) Aim of post-releases monitoring...23 (g) Duration of post-releases monitoring (h) Conclusions of post-release monitoring (i) Results of the release in respect to any risk to human health and the environment...24 E. INFORMATION RELATING TO THE MONITORING PLAN - IDENTIFIED TRAITS, CHARACTERISTICS AND UNCERTAINTIES RELATED TO THE GMO OR ITS INTERACTION WITH THE ENVIRONMENT THAT SHOULD BE ADDRESSED IN THE POST COMMERCIALISATION MONITORING PLAN List of Tables Table :Elements of the transformation fragment PV-ZMIR I3L Table Summary ofn/on 863 Cry3Bbl, NPTII and Cryl Ab protein levels measured in MON 863 and MON 863 x MON 8 10 maize grains Table First year of experiment List of Figures Figure Map of the transformation fragment PV-ZMIR I 3L...14 SNIF 4 jrom insect-protected maize line MON 863 and maize hybrid MON 863 xa1on 8 10

5 SUMMARY NOTIFICATION INFORMATION FORMAT (SNIF) FOR PRODUCTS CONTAINING GENETICALLY MODIFIED HIGHER PLANTS (GMHP) 1. Details of notification (a) (b) (c) (d) 2. Notifier A4. GENERAL INFORMATION Member State of notification :Germany Notification number: C/DE/02/9 Name of the product (commercial and other names): This application under Directive 90/220/EEC and anticipating Directive 200 1/ I8/EC is for import into the European Union, and use therein, of grain and grain products from MON 863 and MON 863 x MON 810 maize. The proposed uses of this maize will be the same as for any other maize, but do not include the cultivation of varieties in the E.U. Date of acknowledgement of notification: (a) Name of notifier: Monsanto Company represented by Monsanto Europe S.A. (b) (c) (d) Address of notifier: Monsanto Europe S.A. Monsanto Avenue de Tervuren 800 N. Lindbergh Boulevard B-i 150 Brussels St. Louis, Missouri BELGIUM USA Is the notifier : domestic manufacturer : [XI importer: [ ] In case of import the name and address of the manufacturer should be given Not relevant 3. General description of the product (a) Name of the recipient or parental plant and the intended function of the genetic modification To produce MON 863 maize, a DNA sequence encoding i) a Cry3Bbl protein variant, MON 863) Cry3Bbl protein, which confers protection to certain coleopteran insects and ii) the NPTII protein (neomycin phosphotransferase II) which provides resistance towards kanamycin for maize plant cell selection purposes, was inserted into maize cells by particle bombardment. For MON 863 x MON 81 0 maize, conventional maize breeding methods have been used to cross the progeny of MON 810 maize (obtained by the introduction of a DNA sequence encoding a Cryl Ab protein, which confers protection to certain lepidopteran insects into maize cells) with the progeny of MON 863 maize. SNIF 5 Placing on the market of maize grilus derived from insect-protected maize line A/ON 863 and maize hybrid MON 863 x MON 810

6 (b) (c) (d) (e) (f) (g) Any specific form in which the product must not be placed on the market (seeds, cut-flowers, vegetative parts, etc.) as a proposed condition of the authorisation applied for This application includes the importation in the European Union and use therein of grain and grain products from MON 863 and MON 863 x MON 8 10 maize and excludes maize varieties and forage derived from MON 863 and MON 863 x MON 810 maize. Therefore, varieties from MON 863 and MON 863 x MON 8 10 maize must not be placed on the market. Intended use of the product and types of users There are no specific differences when MON 863 and MON 863 x MON 810 maize are compared to conventional maize except for their protection, respectively, against certain Coleoptera and Coleoptera/Lepidoptera. MON 863 and MON 863 x MON 81 0 maize have been shown to be substantially equivalent, with exception of the introduced traits, to maize currently in commerce and therefore, the proposed uses and the types of users for grain and maize products derived from MON 863 and MON 863 x MON 810 maize are identical to those for grain and maize products derived from conventional maize. Any specific instructions and/or recommendations for use, storage and handling, including mandatory restrictions proposed as a condition of the authorisation applied for MON 863 and MON 863 x MON 8 10 maize have been demonstrated to be substantially equivalent to other maize, apart from their protection against certain coleopteran and coleopteranllepidopteran insect pests. No specific instructions or recommendations for use, storage or handling of grain or derived products from this maize are envisaged. If applicable, geographical areas within the EXU. to which the product is intended to be confined under the terms of the authorisation applied for Grain and derived products of MON 863 and MON 863 x MON 810 maize are intended for use throughout the E.U. for the purposes specified in the application. Any type of environment to which the product is unsuited This application is for import into the E.U., and use therein, of grain and grain products from MON 863 and MON 863 x MON 81 0 maize and is for use as any other maize grain. Any proposed packaging requirements MON 863 and MON 863 x MON 810 maize have been shown to be substantially equivalent to other maize varieties. Therefore, MON 863 and MON 863 x MON 8 10 maize grain or derived products will be used in the same manner as with other maize and no specific packaging is foreseen. SANIF 6 fromn insect-protected maize line MON 863 and maize hybrid MON 863 x MION 81 0

7 (h) Any proposed labelling requirements in addition to those required by law The grain and derived products of MON 863 and MON 863 x MON 810 maize could be imported into the European Union from a number of' world areas, including Africa, The Americas, Asia and Central and Eastern Europe. In accordance with the requirements of Directive 2001/18/EC, repealing Directive 90/220/EEC on 17 October 2002, Monsanto will undertake a number of measures to ensure that international traders are provided with the necessary information to comply with statutory requirements relating to the placing on the market of MON 863 and MON 863 x MON 810 maize grain. In particular, Monsanto will: a) Informn European and International traders of the approval for import into the European Union of MON 863 and MON 863 x MON 810 maize grain that this product is a genetically modified organism and that MON 863and MON 863 x MON 810 maize grain may be present in bulk shipments of maize grain. To that effect the words "Contains genetically modified organisms" shall appear either on a label or in an accompanying document to the maize grain shipment b) Provide all traders with the commercial name of the product, any agreed European and/or international unique identifier (see question 3.(j)) replacing the current unique code (MON 863, MON 863 x MON 81 0) and any other relevant product information, including procedures for accessing the European public registers of GM organisms c) Advise all traders, and other operators using the product, that MON 863and MON 863 x MON 810 maize grain is subject to the traceability and labelling requirements of Directive 2001/18/1EC and to the requirements of any Community legislation adopted to regulate the traceability and labelling of GM organisms. It should be noted that other genetically modified maize products have been approved for import into the European Union and that MON 863 and MON 863 x MON 810 maize may be marketed in the European Union in mixture with these previously approved maize products. (i) Estimated potential demand i)in the Community In 2001, total production of maize within the fifteen countries comprising the E.U. was estimated at million tonnes. The largest producers being France and Italy. However, the production of MON 863 and MON 863 x MON 81 0 maize is not intended in the E.U. Imports of maize from outside the E.U. including maize grain is about 2.5 million tonnes. (0in export markets for EC supplies Not relevant SNIF 7 Placing on the market Of maize grains derived.from imnsect-protected maize line MON 863 and maize hybrid MON 863 x AMfON 81 0

8 () Unique identification code(s) of the GMIO(s) This application under Directive 90/220/EEC, anticipating Directive 2001/18/EC is for import into the European Union and use therein MON 863 and MON 810 x MON 863 maize, the latter being produced by traditional breeding of two genetically modified inbred lines. one derived from MON 810 event and the other one derived from MON 863 event. The unique identifiers for MON 863, MON and MON 810, MON have been attributed based on the guidance for the designation of a unique identifier for transgenic plants developed by OECD's Working Group on Harnmonisation of Regulatory Oversight in Biotechnology. For MON 863 x MON 810, a combination of those two unique identifiers will be used. 4. Has the GMHP referred to in this product been notified under part B of Directive 2001/1 8/EC and/or Directive 90/220/EEC? Yes I 1 No [X I If no, refer to risk analysis data on the basis of the elements of Part B of Directive 2001/1 8/EC See following sections. 5. Is the product being simultaneously notified to another Member State? Yes I I No [ XI If no, refer to risk analysis data on the basis of the elements of Part B of Directive 200 1/18/EC See following sections. OR Has the product being notified in a third country either previously or simultaneously? Yes [XI No HI If yes, please specify Canada, Japan and USA 6. Has the same GMHP been previously notified for marketing in the Community? Yes [I No [ X 7. Measures to take in case of unintended release or misuse as well as measures for disposal and treatment Maize is not an invasive plant because it is a weak competitor outside cultivation. For this reason, volunteer maize is not found in non-crop situations, for example, in fence or hedgerows, ditches, and roadsides. It is highly unlikely, therefore, that any grain disseminated into the environment would pose any threat to the environment. In the unlikely event of establishment, volunteer plants could be easily controlled by currently available selective herbicides (e.g. fluazifop-p-butyl) or by mechanical SNIF from insect-protected maize line MO0N 863 and maize hybrid MON 863 x MON 81 0

9 means. Therefore no specific measures are recommended in case of unintended release of MON 863 and MON 863 x MON 810 maize. Misuse of the imported grain as seed remains extremely unlikely since 1) only crified hybrid maize seed can be marketed in the E.U. and 2) the (F2) grain, coming from open-pollinated production fields, will not have the agronomic yield potential and homogeneity of true FlI hybrid seed. The measures for waste disposal and treatment for MON 863 and MON 863 x MON 8 10 maize products are the same as those for other maize products. B. NAATURE OF THE GMHP CONTAINED IN THE PRODUCT INFORMATION RELATING TO THE RECIPIENT OR (WHERE APPROPRIATE) PARENTAL PLANTS 8. Complete name (a) (b) Family name: Gramineae Genus :Zea (c) Species m nays (2n = 20) (d) (e) (f) Subspecies :Not applicable Cultivar/breeding line :specifically, the maize plant tissue that was the recipient of the introduced DNA was a cell culture designated AT824, initiated from immature embryos of an inbred maize line (AT) Common name: Maize; Corn 9. (a) Information concerning reproduction ()M~ode(s) of reproduction Maize (Zea inays) reproduces sexually, is a wind-pollinated, monoecious species with separate stamninate (tassels) and pistillate (silk) flowers, which encourages natural pollination between maize plants. Wind movements across the maize field cause pollen from the tassel to fall on the silks of the same or adjoining plants. Self-pollination leads to homogeneity of the genetic characteristics within a single plant while cross-pollination combines the genetic traits of many plants. (ii} Specific factors affecting reproduction, if any Tasselling, silking, and pollination are the most critical stages of maize development and, consequently, grain yield may ultimately be greatly impacted by moisture and fertility stress. Under conditions of high temperature and desiccation, maize pollen viability is measured in minutes; these conditions may even destroy the tassel before any viable pollen is shed. More moderate conditions can extend the field life of pollen to hours. SNIF 9 fromn insect-protected maize line MON 863 and maize hybrid MON 863 x MVON 81 0

10 (b) (iii) Generation time Maize is an annual crop with a cultural cycle ranging from as short as 60 to 70 days to as long as 43 to 48 weeks from seedling emergence to maturity. Sexual compatibility with other cultivated or wild plant species Out-crossing with cultivated Zea varieties Maize is wind pollinated, and the distance that viable pollen can travel depends on prevailing wind patterns, humidity, and temperature. All maize will interpollinate, except for certain popcorn varieties and hybrids that have one of the gametophyte factors (Gas, Ga, and ga allelic series on chromosome 4). Maize pollen, therefore, moves freely within an area, lands on silks of the same variety or different varieties, germninates almost immediately after pollination, and within 24 hours completes fertilisation. Out-crossing with wild Zea species Annual teosinte (Zea mays ssp. Mexicana, formnerly Euchlaena mexicana) (2n 20) and maize (Zea mays L.) (2n = 20) are wind pollinated, tend to out-cross, and are highly variable, interfertile subspecies. A frequency of one Fl hybrid (maize x teosinte) for every 500 maize plants or 20 to 50 teosinte plants in the Chalco region of the Valley of Mexico wvas reported. Out-crossing and gene exchange between teosinte and maize occur freely, and, accompanied by selection, teosinte had a significant role in the evolution of maize. Teosinte, however, is not present in either Europe or the U.S. "Corn Belt". The natural distribution of teosinte is limited to the seasonally dry, subtropical zone with summer rain along the western escarpment of Mexico and Guatemala and the Central Plateau of Mexico. Tripsacum species are perennials and seem to be more closely related to the genus Man isuris than to either maize or teosinte. Special techniques are required to hybridise maize and Tripsacumn. Except for Tripsacum floridanum, it is difficult to cross Tripsacum with maize, and the offspring of the cross show varying levels of sterility. Tripsacum-maize hybrids have not been observed in the field and Tripsacumteosinte hybrids have not been produced. 10. Survivability (a) (b) Ability to form structures for survival or dormancy Maize is an annual crop and seeds are the only survival structures. Natural regeneration from vegetative tissue is not known to occur. Specific factors affecting survivability, if any Maize cannot survive without human assistance and is not capable of surviving as a weed due to past selection in its evolution. The appearance of maize in rotational fields following the maize crop from the previous year is rare under European conditions. Maize volunteers are killed by frost or easily controlled by current agronomic practices including cultivation and the use of selective SNIF 10 fromn inscct-protected maize line MON 863 and maize hybrid MON 863 x MON 81 0

11 11. Dissemination herbicides. Temperatures above 45 0 C have also been reported as injurious to maize seed viability. (a) (b) Ways and extent of dissemination Dissemination may occur by means of seed and pollen dispersal. Although pollen might be spread over long distances, maize pollen viability is highly dependent on temperature and desiccation. Specific factors affecting dissemination, if any In contrast to weedy plants, maize has a polystichous female inflorescence (ear) on a stiff central spike (cob) enclosed in husks (modified leaves). Consequently, seed dispersal of individual kernels does not occur naturally because of the structure of the ears of maize. Seed dissemination is impacted by mechanical harvesting and transport as well as insect or wind damage, all of which may cause some mature ears to fall to the ground, where they could remain un-harvested. Genetic material can be disseminated by pollen movement. Pollen dispersal is influenced by wind and weather conditions. Measuring about 0.1I mm in diameter, maize pollen is the largest of any pollen normally disseminated by wind from a comparably low level of elevation. Dispersal of maize pollen is influenced by its large size and rapid settling rate. 12. Geograph ical distribution of th e planti Maize, because of its many divergent types, is grown over a wide range of climatic conditions. The bulk of the maize is produced between latitudes 300 and 550, with relatively little grown at latitudes higher than 470 latitude anywhere in the world. The greatest maize production occurs where the warmest month isothenrms range between 21 and 270 C and the freeze-free season lasts 120 to 180 days. A summer rainfall of 15 cm is approximately the lower limit for maize production without irrigation with no upper limit of rainfall for growing maize, although excess rainfall will decrease yields and increase the chance of fungal infection. 13. In the case of plant species not normally grown in the Member State(s), description of the natural habitat of the plant, including information on natural predators, parasites, competitors and symbionts Maize is widely grown in the European Union and represents a significant portion of global maize production. Significant areas of maize production in Europe include the Danube Basin from southwest Germany to the Black Sea along with southern France through the Po Valley of nor-thern Italy. 14. Potentially significant interactions of the plant with other organisms in the ecosystem where it is usually grown, including information on toxic effects on humans, animals and other organisms There are no known toxic effects of the maize plant to humans, animals or livestock; it has a history of safe use for human food and animal feed. However, maize is known to interact with other organisms in the environment including insects, birds, and mammals. It is susceptible to a range of fungal diseases and nematode, insect and mite pests. SNIF 1 Placing on the market of maize gra ins derived from insect-protected maize line MON 863 and maize hybrid MON 863 x MON 810

12 15. Phenotypic and genetic traits Maize (Zen mavys) is a wind-pollinated, monoecious species with separate staminate (tassels) and pistillate (silk) flowers, which encourages the natural out-crossing between maize plants. Self-pollination leads to homogeneity of the genetic characteristics within a single plant while cross-pollination combines the genetic traits of many plants. This inbred-hybrid concept and resulting yield response is the basis of the modem seed maize industry. INFORMATION RELATING TO THE GENETIC MODIFICATION 16. Description of the methods used for the genetic modification MON 863 and MON 81 0 maize were modified by incorporation of a DNA fragment derived, respectively, from plasmid vectors PV-ZMIiR1 3 and PV-ZMBKO7 into the maize genome using a particle acceleration method. MON 863 x MON 810 maize has been obtained by conventional breeding of MON 863 and MON 81 0 maize lines. 17. Nature and source of the vector used The vector used to amplify the DNA fragment which was introduced in MON 863 is composed of a puc plasmid replication origin associated with a selectable marker, nptlf. The functions carried by the vector are required to allow its maintenance and amplification in E coli bacterial cells. Like the original puc vectors, this vector does not contain transfer origins, i.e., sequences allowing transfer from bacteria to bacteria. The vector used for the genetic modification of MON 81 0 maize was described in the application pursuant to Council Directive 90/220/EEC [Commission Decision of 22 April 1998 concerning the placing on the market of genetically modified maize (Zen ma ys L. line MON 810), pursuant to Council Directive 90/220/EEC (98/294/EC - Official Journal of the European Communities, L- 13 1/32 of May 05, 1 998)]. MON 863 x MON 810 maize has been obtained by conventional breeding of MON 810 and MON 863 maize lines and no vector has been used to produce this maize hybrid. 18. Size, source [aine of donor organism(s)] and intended function of each constituent fragment of the region intended for insertion The transformation fragment PV-ZMTR1 3L contained the two genes to be introduced in the plant cells, i.e., the chimearic cry3bb] gene (encoding the agronomic trait) and the npt~l gene (selectable marker) designed to be expressed in the maize cells. The expression cassettes (see Table and Figure 18.1.) corresponding to these two genes consist of respectively: i) the nptl ORF regulated by the 35S promoter, and the NOS 3' polyadenylation sequence and ii) a cry3bbl open reading frame (ORF) regulated by the 4-AS1I plant promoter and the wtcab leader, rice actin intron and tahspl17 3' polyadenylation sequence. SNIF 12 fr~om insect-protected maize line MON 863 and maize hybrid MON 863 xamon 810

13 Table :Elements of the transformation fragment PV-ZMIRI3L Sequence Size (Kb) Source Function MON 863 crv3bbl gene cassette 4ASI 0.22 Cauliflower Mosaic Virus Promoter (CMV) wt CAB 0.06 Wheat (Triticum aestivum) Enhances translation racti intron 0.49 Rice (Oryza sativa) Enhances transcription MON 863 criv3bbl Bacillus thuringiensis subsp. ORE encoding kumamotoensis resistance to corn rootworm tahsp 17 3' 0.23 Wheat (Triticurn aestivum) Ends transcription and directs polyadenylation Selectable marker elements 35 S 0.32 Cauliflower Mosaic Virus Promoter (CMV) nptlj 0.82 E. coli ORF allowing plant cell selection NOS 3' 0.26 Agrobacteriurn tuineftiiens Ends transcription and directs polyadenylation SNIF 13 f/rom insect-protected maize line MON 863 and maize hybrid MON 863 x MON 810

14 Figure :Map of the transformation fragment PV-ZMIR13L HindIll 12 Ncol 1352 EcoRi 4417 EcoRI 1358 FEcoRi 4673 EcoRV 1624 Hindill S promoter L.J ~500 nucleotides nptli ORF NOS 3' (ends transcription and directs polyadenylation) 4AS I promoter wt CAB - wheat chlorophyll a/b-binding protein [I (translation enhancement) racti intron (transcription enhancement) W M0N863 crvi3bbl ORF tahsp wheat heat shock protein ii&~(ends transcription and directs polyadenylation) Inform-ation related to MON 810 maize is sumnmarised in the Commission Decision of 22 April 1998 concerning the placing on the market of genetically modified maize (Zea mavys L. line MON 810), pursuant to Council Directive 90/220/EEC (98/294/EC- Official Journal of the European Communities, L 13 1/32 of May 05, 1998). MON 863 x MON 810 maize has been obtained by conventional breeding of MON 810 and MON 863 maize lines and no additional vector has been used to produce this maize hybrid (see transform-ation of parental lines). INFORMATION RELATING THE GMHP 19. Description of the trait(s) and characteristics which have been introduced or modified MON 863 maize plants are protected from damage due to feeding by CRW larvae. The tissues of MON 863 maize contain a modified cry3bbl gene, derived from Bacillus thuringiensis, which encodes a modified B.t. Cry3Bbl protein for CRW control. SNIF 14 Placing on the market oj'maize grains derived /rom insect-protected maize line MO0N 863 and maize hybrid MON 863 x. MON 81 0

15 MON 863 maize that expresses the modified Cry3Bbl protein from B.t. offers an entirely new means to control CRW that is safe for humans and the environment. As the leading pest of U.S. maize, based on insecticide use, there is an ongoing need for efficacious CRW control measures. CRW-protected maize will offer growers a unique new management tool for CRW, which will reduce or eliminate the risks associated with chemical transportation, storage, application, disposal, and stewardship. Agro-ecosystems will benefit from the specificity of the product to CRW and the lack of harmful effects on beneficial insects or wildlife. CRW-protected maize is fully compatible with current management protocols for CRW, including integrated pest management (1PM). The Bit. protein, MON 863 Cry3Bbl has shown high levels of activity against the CRW complex. MON 863 maize demontrates levels of control that are superior to those of commercial soil-applied insecticides. MON 863 x MON 810 maize consists of varieties developed using conventional methods of maize breeding which express: 1. the Cryl Ab protein, derived from Bacillus thuringiensis subsp. kurstaki, which prvdes protection from certain Lepidopteran insect pests (including Ostrinia nubilalis (European corn borer) and Sesamia spp), 2. the modified Cry3Bbl protein, derived from Bacillus thuringiensis subsp. kumamotoensis, which provides protection from certain Coleopteran pests (corn rootwormn). 20. Information on the sequences actually inserted/deleted/modified (a) Size and structure of the insert and methods used for its characterisation, including information on any parts of the vector introduced in the GMHP or any carrier or foreign DNA remaining in the GMHP MON 863 maize was produced by particle acceleration technology using a MWlul DNA restriction fragment from plasmid PV-ZMIRl1 3 containing the nptll and MON 863 crv3bbl cassettes. The MON 863 event contains one DNA insert located on a 5.0 Kb Nd/el fragment. This insert contains one copy of the fragment used in transformation. No additional elements from the DNA fragment used in transformation, linked or unlinked to intact cassettes, were detected in the genome. PCR and DNA sequencing were used to verify the 5' and 3' junction sequences of the insert with the plant genome, as well as the intactness of the 5' and 3' ends of the insert. Approximately 10 bp from the 3' end of PV-ZMIR13L, including the Hint/lll restriction site, is missing; however, the tahspl7 3' polyadeylation sequence is intact. Additionally, event MON 863 does not contain any detectable plasmid backbone sequence including ori-puc or the nptii coding region regulated by a bacterial promoter. These data support the conclusion that only the two expected full length proteins, MON 863 Cry3Bbl and NPTII, should be encoded by the insert in event MON 863. Molecular analysis related to MON 810 maize is summarise in the Commission Decision of 22 April 1998 concerning the placing on the market of genetically modified maize (Zea mays L. line MON 810), pursuant to Council Directive 90/220O/EEC (98 '294/EC- Official Journal of the European Communities, L 13 1/32 of May 05, 1998). SNIF 15 firom insect-protected maize line MO0N 863 and maize hybrid MON 863 x MON 81 0

16 (b) (c) (d) (e) The genome of MON 863 x MON 810 maize contains two different inserts, one derived from MON 810 maize and one derived from MON 863 maize. Southern blot analyses demonstrated that the size and structure of each insert are identical to those of their respective parents and therefore that their molecular structure has been preserved during the breeding process In case of deletion, size and function of the deleted region(s) Not applicable Location of the insert in the plant cells (integrated in the chromosome, chloroplasts, mitochondria, or maintained in a non-integrated form), and methods for its determination Southern blot analysis was conducted to confirm the location and stability of the inserted DNA in MON 863 maize. One single copy of the transform-ation fragment has been inserted at one site of the plant nuclear genome. This copy contains two ORFs, MON 863 cry3bbl and njptii, which are expressed. Segregation of the traits occurs according to Mendelian genetics. The integration of MON 810 insert in the nuclear genome has been previously established [Commission Decision of 22 April 1998 concerning the placing on the market of genetically modified maize (Zea may-s L. line MON 810), pursuant to Council Directive 90/220/EEC (98/294/EC- Official Journal of the European Communities, L 13 1/32 of May 05, 1998)]. MON 863 x MON 810 maize results from the conventional breeding of MON 810 and MON 863 maize and thus location of inserts has not been altered during the breeding process. Copy number and genetic stability of the insert MON 810 maize and MON 863 maize each contain a single insert at one locus in the maize genorne. Therefore, the MON 863 x MON 8 10 product contains two distinct simple inserts. In case of modifications other than insertion or deletion, describe function of the modified genetic material before and after the modification as well as direct changes in expression of genes as a result of the modification Not applicable 21. Information on the expression of the insert (a) Information on the expression of the insert and methods used for its characterisation Levels of MON 863 Cry3B~bl, NPTII and CrylAb proteins were determnined in tissues collected from MON 863 and MON 863 x MON 8 10 maize plants grown under field conditions at multiple sites, representative of regions where corn rootworm protected maize would be grown as commercial products. Direct double antibody sandwich enzyme-linked immunosorbent assay (ELISA) methods wvere developed and validated to quantify the levels of MON 863 Cry3Bbl, NPT11 and CrylAb proteins in tissue extracts of MON 863 and MON 863 x MON 810 maize and control plants. MON 863 Cry3BblI, NPTII SNIF 16 fr-om insect-protected maize line MO0N 863 and maize hybrid MON 863 x MON 81 0

17 and CrylAb levels were below the limit of detection in control plant tissues and thus, are not reported. (b) Parts of the plant where the insert is expressed (e.g. roots, stem, pollen, etc.) This application being restricted to import of grain and derived products from MON 863 and MON 863 x MON 810 maize, the only relevant part of the plant for protein expression levels is the grain. Table summarizes the level of proteins in grain samples. Table 21.1.: Summary of MON 863 Cry3Bbl, NPTII and CrylAb protein levels measured in MON 863 and MON 863 x MON 810 maize grains [average (range) in ~tg/g fw] MON 863 maize MON 863 x MON_810 maize MON 863 Cry3BbI NPTII MON 863 Crv3Bbl -- NPTII _CrvlAb 70 (49-86) < LOD 42.7 (<LOD'-84.1) < LOD* 61.1 ( ) < LOD' 0.84 ( ) Below the limit of detection ptglg fw Below the limit of detection :0.096 ~.tg/g fw First and second lanes correspond to field trials in the US (1999) and Argentina ( ), respectively. 22. Information on how the GMHP differs from the recipient plant in (a) Mode(s) and/or rate of reproduction Agronomic equivalency (seedling vigour; early stand count; GDU' to 50% pollen shed; GDLi to 50% silk emergence; ear height; plant height; final stand count; test weight; grain moisture; yield) has been evaluated in multiple field trials with six MON 863 hybrids. Statistically significant differences were observed for very few of the parameters evaluated. These differences were uniform-tly small, not consistently observed across locations or other MON 863 hybrids, and none would be considered to be of adverse agronomic consequence. The results of these trials support a conclusion of agronomic equivalence for MON 863 maize and their respective controls. Information related to MON 810 maize was described in the application pursuant to Council Directive 90/220/EEC [Commission Decision of 22 April 1998 concerning the placing on the market of genetically modified maize (Zea mays L. line MON 810), pursuant to Council Directive 90/220/EEC (98/294/EC- Official Journal of the European Communities, L 1-3)1/32 of May 05, 1998)]. Growing degree units (GDU) = f(tmax + Tmin)12] -- 50; where Tmnax 31 0 C and Tmnin Ž? 13'C SNIF 17 firom insect-protected maize line MON 863 and maize hybrid MON 863 x MO0N 81 0

18 (b) (c) Based on the conclusion established for each parent, no differences are anticipated in the reproductive capability of MON 863 x MON 810 maize when compared to the parental lines and therefore MON 863 x MON 810 maize should behave similarly to conventional lines. Dissemination This application being restricted to import of maize grain, dissemination of flowering maize plants in Europe is likely to be limited. For a cross pollination to occur between MON 863 derived plants and other maize plants, release would have to take place within proximity of other maize plants. Based on the above-mentioned results, the mode of reproduction of MON 863 maize is equivalent to that of conventional maize and thus its dissemination capabilities (by seed or pollen) will also be equivalent. Outcrossing with wild Zea species Outerossing to wild Zea species will not occur as there are no wild related species of maize in Europe (see Section B.2.). Outcrossing to cultivated maize varieties Gene exchange between MON 863 maize and cultivated conventional and genetically modified maize will be similar to natural crossing between conventional varieties (see Section B.2.). Wind blown pollen would move among plants within the same field and among plants in nearby fields at low frequencies. If outcrossing occurs, the production of MON 863 Cry3Bbl protein in maize would not raise safety concerns due to the demonstrated safety of the protein to humans. Informnation related to MON 810 maize was described in the application pursuant to Council Directive 90/220/EEC [Commission Decision of 22 April 1998 concerning the placing on the market of genetically modified maize (Zea mays L. line MON 810), pursuant to Council Directive 90/220/EEC (98/294/EC- Official Journal of the European Communities, L 13 1/32 of May 05, 1998)1. Based on the conclusion established for each parent, no differences are anticipated in dissemination of MON 863 x MON 8 10 maize when compared to the parental lines and therefore MON 863 x MON 81 0 maize should behave similarly to conventional lins Survivability This application being restricted to import of maize grain, release of viable seeds in Europe is likely to be limited. Analysis of the agronomic parameters from MON 863 maize have shown no difference with non-transgenic control hybrids and thus MON 863 maize is not expected to behave differently from conventional maize with respect to survivability. To assess whether MON 863 maize could have a selective advantage, with respect to pest infestation, a maize line containing the MON 863 event was compared to the non-transgenic control hybrid for susceptibility to a series of pests and diseases. SNIF 18 P/acing on the market Of maize grains derived from insect-protected maize line MON 863 and maize hybrid MON 863 x MON 81 0

19 (d) Apart from protection against CRW, no differences in disease severity or insect infestation were detected between MON 863 and non-transgenic control plants. The only new genetic trait specific to the line containing MON 863 event is the insecticidal property carried by the inserted DNA sequence and directed against the coleopteran pest of maize corn rootworm (CRW, Diabrotica sp). Weediness potential of MON 863 maize Modern maize cannot survive as a weed because of the past selection for agronomic criteria which guided the evolution of maize. In contrast to weedy plants, the maize car is enclosed within husks. Consequently, seed dispersal of individual kernels does not occur naturally because of this structure. Even if imported maize grain was to be more widely disseminated, volunteer maize is not found growing in fence rows, ditches, or road side as a weed. Maize is poorly suited to survive without human assistance and is not capable of surviving as a weed. Maize lines derived from MON 863 event can be considered substantially equivalent to their non-transgenic counterparts except for the presence of the introduced traits. The presence of these genes is unlikely to increase the weediness potential of MON 863 maize. Maize lines derived from MON 863 event are thus expected to behave like their non-transgenic counterparts and thus not to be able to survive as weeds. Information related to MON 81 0 maize was described in the application pursuant to Council Directive 90/220/EEC [Commission Decision of 22 April 1998 concerning the placing on the market of genetically modified maize (Zea mays L. line MON 8 10), pursuant to Council Directive 90/220/EEC (98/294/EC- Official Journal of the European Communities, L 131/32 of May 05, 1998)]. Based on the conclusion established for each parent, no differences are anticipated in survivability of MON 863 x MON 8 10 maize when compared to the parental lines and therefore MON 863 x MON 81 0 maize should behave similarly to conventional lines. The scope of the use of MON 863 x MON 810 maize in the European Union excludes cultivation of varieties. Other differences No other differences observed. 23. Potential for transfer ofgenetic material from the GMHP to other organisms Based on the fact that pollen production and pollen viability as measured by yield and germination of progeny are unchanged by the genetic modification, the outcrossing frequency to other maize varieties or to wild relatives (which are not present in the E.U.) is unlikely to be different for MON 863 and MON 863 x MON 810 maize when compared to other varieties. Also, none of the genetic elements introduced in the plant carries genetic transfer function. Furthermore neither MON 863 nor MON 863 x MON 81 0 cultivation is within the scope of this application. SNIF 19 Jrom inseet-protectedl maize line MON 863 and maize hybrid MON 863 x MON 81 0

20 24. Information on any harmful effects on human health and the environment, arising from the genetic modification An assessment of the human safety of the MON 863 Cry3Bbl, NPTII and CrylAb proteins was conducted based upon the extensive characterization of those proteins. The human safety of the MON 863 Cry3Bbl, NPTII and CrylAb proteins has been established based upon the following considerations: (I) no amino acid sequence simlilarity to known toxins, other than B.t. proteins inthe case of MON 863 Cry3Bbl and CrvlAb, and no immunologically relevant sequence similarity with known allergens, (2) rapid degradation under conditions v~hich simulate mammalian digestive systems, (3) no indications of acute toxicity in mice administered MON 863 Cry3Bbl, NPTII or CrylAb protein by oral gavage, (4) very low dietary exposure, and (5) a history of safe use. Finally, the nutritional equivalence of grain from MON 863 and MON 863 x MON 810 mnaize has been established by compositional analysis and the wholesomeness of MON 863 and MON 81 0 maize grain, confirmed by feeding to broiler chickens. 25. Information on the safety of the GMHP to animal health, where the GMHP is intended to be used in animal feedstuffs, if different from that of the recipient/parental organism(s) There is no difference between MON 863, MON 863 x MON 81 0 maize and the recipient organism in terms of safety to animals. An assessment of the safety of the MON 863 Cry3Bbl, NPT1I and CrylAb proteins was conducted based upon the extensive characterization of those proteins. The safety of the MON 863 Cry3Bbl, NPT1I and CrylAb proteins has been established based upon the following considerations: (1) no amino acid sequence simi'larity to known toxins, other than B.t. proteins inthe case of MON 863 Cry3Bbl and Cry lab, and no immunologically relevant sequence similarity with known allergens, (2) rapid degradation under conditions which simulate mammalian digestive systems, (3) no indications of acute toxicity in mice administered MON 863 Cry3Bbl, NP1TII or CrylAb protein by oral gavage, (4) very low dietary exposure, and (5) a history of safe use. The Cry3lBb and CrylAb proteins are present in commercially available insecticides, which are considered environmentally acceptable, because they are specific for the target insect pest and are typically harmless to plants and other non-target organisms. CrylI Ab maize has been grown commercially on 20.5 million hectares since 1997 and has been widely used in the E.U. and other countries as a source of feed ingredients. Finally, the nutritional equivalence of grain and forage from MON 863 and MON 863 x MON 810 maize has been established by compositional analysis and the wholesomeness of MON 863 and MON 810 maize grain, confirmned by feeding to broiler chickens. SNIF 20 Placing on the market Of maize grains derived from insect-protected maize line MON 863 and maize hybrid MO0N 863 x MON 810

21 26. Mechanism of interaction between the GMHP and target organisms (if applicable), if different from that of the recipient/parental organism(s) Not relevant to the current application since this application is restricted to import of grain and grain derived products from MON 863 and MON 863 x MON 81 0 maize and excludes E.U. cultix~ation of maize varieties expressing MON 863 Cry3Bbl Potentially significant interactions with non-target organisms, if different from the recipient or parental organism(s) Not relevant to the current application since this application is restricted to import of grain and grain derived products from MON 863 and MON 863 x MON 810 maize and excludes E.U. cultivation of MON 863 and MON 863 x MON 810 maize varieties. 28. Description of detection and identification techniques for the GMHP, to distinguish it from the recipient or parental organism(s) Southern blot or PCR techniques may be employed for the detection and identification of the inserted nucleotide sequences. Specific ELISAs have been developed and could be used to detect MON 863 Cry3BbI1, NPTII or Cryl Ab proteins in individual plants and their grains. A detection method specific for MON 863 transform-ation event was provided to the Genrnan Competent Authority in charge of the evaluation of this application. In the case of MON 810 maize, which is authorised under Directive 90/220/EEC and Regulation 258/97, a validation process is currently being organised through a collaboration with AACC (American Association of Cereal Chemists), BgVV, GeneScan and JRC. The BgVV (Germany) has also developed an event-specific method for MON 810 ["Commission Recommendation of 25 January 2002 concerning a coordinated programme for the official control of foodstuffs for 2002" (2002/66/EC - Official Journal of the European Communities, L 26/8 of January 30, 2002)]. MON 810 x MON 863 maize will be detectable using either the event-specific PCR method for detecting the introduced DNA present in the parental lines, MON 810 maize or the equivalent method for MON 863 maize. INFORMATION ON THE POTENTIAL ENVIRONMENTAL IMPACT FROM THE RELEASE OF THE GMHP 29. Potential environmental impact from the release or the placing on the market of GMOs (Annex II, D2 of Directive 2001/118/FEC, if different from a similar release or placing on the market of the recipient or parental organism(s) Not relevant to the current application since this application is restricted to import of grain and grain derived products and excludes E.U. cultivation of MON 863 and MON 863 x MON 810 maize varieties. SNIF 21 from insect-protected maize line MON 863 and maize hybrid MON 863 x MON 810